scholarly journals Relationship Between Protein Synthesis and Secretion in Liver Cells and the State of the Adenine Nucleotide System

1979 ◽  
Vol 32 (3) ◽  
pp. 299 ◽  
Author(s):  
Kaylene Edwards ◽  
Jörg Urban ◽  
Gerhard Schreiber
Keyword(s):  
Protein Synthesis ◽  
Oxidative Phosphorylation ◽  
Protein Secretion ◽  
Liver Cell ◽  
Maximum Rate ◽  
Adenine Nucleotide ◽  
Energy Charge ◽  
Adenylate Energy Charge ◽  
Atp Level ◽  
Atp Pool

Adenine nucleotide levels could be precisely and reproducibly adjusted in liver cell suspensions by partially depleting the ATP pool with D-fructose or glycerol. Thus, it was possible to quantitatively correlate rates of protein synthesis and secretion with intracellular levels of ATP and with derived parameters, such as the adenylate energy charge. Half the maximum rate of incorporation of leucine into protein was observed at an energy charge of 0�80, a ratio of ATP to ADP of 2�6, and an ATP level of 1�05 pmol per g of wet cells. Proteins were secreted with half the maximum rate at an energy charge of 0�85, a ratio of ATP to ADP of 3�1 and an ATP concentration of 1 �1 pmol per gof wet cells. Protein secretion dill not depend on continued synthesis. Inhibitors of oxidative phosphorylation inhibited protein secretion in addition to protein synthesis, in contrast to observations by other authors on liver slices.

scholarly journals Labeling of the releasable adenine nucleotides of washed human platelets

Blood ◽  
1977 ◽  
Vol 49 (1) ◽  
pp. 89-99 ◽  
Author(s):  
HJ Reimers ◽  
MA Packham ◽  
JF Mustard
Keyword(s):  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Energy Charge ◽  
Human Platelets ◽  
Transport Processes ◽  
Adenylate Energy Charge ◽  
Prolonged Incubation ◽  
Atp Pool ◽  
Adenine Nucleotide Pool

Abstract In rabbit platelets, the metabolically active ATP pool equilibrates with the releasable ATP pool within 1 day. The studies showing this have now been extended to human platelets. Human platelets labeled with 14C-adenosine or 14C-adenine were incubated for up to 10 hr in vitro at 37 degrees C. After 10 hr, about 12% of the total platelet 14C-ATP and 14C-ADP had become releasable with thrombin (4.2 units/ml). Lysis of platelets did not occur, since less than 1% of the platelet-bound 51Cr from platelets labeled with this radioisotope appeared in the ambient fluid upon thrombin treatment. The 14C-ATP/14C-ADP ratio of the released adenine nucleotides (7.6) was similar to the 14C-ATP/14C-ADP ratio of the nonreleasable adenine nucleotides (7.1) 2 hr after the labeling with 14C-adenosine. However, upon prolonged incubation (10 hr) in vitro, the 14C-ATP/14C-ADP ratio of the releasable adenine nucleotides decreased to 2.7. The adenylate energy charge and the 14C- ATP/14C-ADP ratio of the metabolic adenine nucleotide pool did not change significantly during the time of observation. The 14C-ATP content of the platelets decreased by less than 1% hr of incubation at 37 degrees C. These observations are interpreted to mean that the 14C is transferred from the metabolically active, nonreleasable adenine nucleotide pool of human platelets into the releasable adenine nucleotide pool as ATP and is partially hydrolyzed there to yield ADP. The transfer of ATP across the storage organelle membrane of platelets may be similar to transport processes in the chromaffin cells of the adrenal medulla and may represent a general phenomenon in cells that possess storage organelles containing adenine nucleotides.

scholarly journals Labeling of the releasable adenine nucleotides of washed human platelets

Blood ◽  
1977 ◽  
Vol 49 (1) ◽  
pp. 89-99 ◽  
Author(s):  
HJ Reimers ◽  
MA Packham ◽  
JF Mustard
Keyword(s):  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Energy Charge ◽  
Human Platelets ◽  
Transport Processes ◽  
Adenylate Energy Charge ◽  
Prolonged Incubation ◽  
Atp Pool ◽  
Adenine Nucleotide Pool

In rabbit platelets, the metabolically active ATP pool equilibrates with the releasable ATP pool within 1 day. The studies showing this have now been extended to human platelets. Human platelets labeled with 14C-adenosine or 14C-adenine were incubated for up to 10 hr in vitro at 37 degrees C. After 10 hr, about 12% of the total platelet 14C-ATP and 14C-ADP had become releasable with thrombin (4.2 units/ml). Lysis of platelets did not occur, since less than 1% of the platelet-bound 51Cr from platelets labeled with this radioisotope appeared in the ambient fluid upon thrombin treatment. The 14C-ATP/14C-ADP ratio of the released adenine nucleotides (7.6) was similar to the 14C-ATP/14C-ADP ratio of the nonreleasable adenine nucleotides (7.1) 2 hr after the labeling with 14C-adenosine. However, upon prolonged incubation (10 hr) in vitro, the 14C-ATP/14C-ADP ratio of the releasable adenine nucleotides decreased to 2.7. The adenylate energy charge and the 14C- ATP/14C-ADP ratio of the metabolic adenine nucleotide pool did not change significantly during the time of observation. The 14C-ATP content of the platelets decreased by less than 1% hr of incubation at 37 degrees C. These observations are interpreted to mean that the 14C is transferred from the metabolically active, nonreleasable adenine nucleotide pool of human platelets into the releasable adenine nucleotide pool as ATP and is partially hydrolyzed there to yield ADP. The transfer of ATP across the storage organelle membrane of platelets may be similar to transport processes in the chromaffin cells of the adrenal medulla and may represent a general phenomenon in cells that possess storage organelles containing adenine nucleotides.

Energy Control During Platelet Secretion:Predominant Role Of ATP-Turnover

1981 ◽  
Author(s):  
A Deijns ◽  
J W N Akkerman
Keyword(s):  
Control Function ◽  
Energy Charge ◽  
Dense Granule ◽  
Metabolic Energy ◽  
Adenylate Energy Charge ◽  
Glucose Addition ◽  
Atp Level ◽  
Atp Turnover ◽  
Dense Granule Secretion ◽  
Granule Secretion

Platelet aggregation and secretion of granular contents require metabolic energy. This implies the existence of a control mechanism that adjusts the rate of energy producing pathways to the energy need of these functions. Such a control function has been attributed to the level of metabolic ATP, to the adenylate energy charge (AEC =(ATP + 1/2ADP)/(ATP + ADP + AMP) and to the velocity of energy generation and consumption (ATP-turnover). In this study we investigated which factor dominates in control of dense granule (3H-serotonin)-, α-granule (β thromboglobulin)- and lysosomal granule (N acetyl β glucosaminidase) secretion. Human gel- filtered platelets were incubated in glucose-free, CN containing medium. Under these conditions ATP-generation only took place in glycogenolysis, which alone was unable to maintain ATP homeostasis. Consequently, metabolic ATP and AEC fell. Addition of glucose restored glycolytic ATP resynthesis wich restored the AEC but the loss of ATP was for the main part irreversible due to hypoxanthin formation. With this system a broad range of metabolic ATP levels and AEC’s could be obtained both at decreasing ATP turnover (before glucose addition) and at increasing ATP turnover (after glucose addition). Analysis of secretion velocity of dense-, α- and lysosomal granules after initiation with thrombin (5 U/ml) showed that at decreasing turnover the secretion velocity of all types of granules depended on both metabolic ATP level and AEC. In contrast, at increasing ATP turnover the correlation between secretion velocity and metabolic ATP level or AEC was lost. Instead, dense granule secretion was faster and lysosomal granule secretion was slower than expected on the basis of metabolic ATP level or AEC, whereas α-granule secretion showed intermediate levels. The data indicate that the three types of granule secretion differ in their dependence on metabolic energy and that apart from metabolic ATP and AEC, ATP turnover is of crucial importance for secretion of granular contents.

Changes in nucleotide pools during conidial germination in Neurospora crassa

1981 ◽  
Vol 59 (11-12) ◽  
pp. 899-905 ◽  
Author(s):  
Richard L. Sabina ◽  
Paulette Dalke ◽  
Alan R. Hanks ◽  
Jane M. Magill ◽  
Clint W. Magill
Keyword(s):  
Neurospora Crassa ◽  
Adenine Nucleotide ◽  
Energy Charge ◽  
Fold Increase ◽  
Conidial Germination ◽  
Adenylate Energy Charge ◽  
Wild Type ◽  
Nucleotide Pools ◽  
Nucleotide Sugars ◽  
Purine Pathway

The acid-soluble nucleotide pools of wild type and several adenine auxotrophs of Neurospora crassa were studied immediately prior to and during conidial germination in the presence of adenine. A two- to four-fold increase in most nucleotide pools was observed after 6 h of germination at 33 °C indicating a general increase in nucleotide pools during this developmental period. The largest components of the acid-soluble nucleotide pools were uracil-containing nucleotide–sugars, which are precursors of chitin and glucan, the major constituents of cell walls. On removal of adenine, the UDP–sugar pools decreased significantly, in adenine auxotrophs, while the pools of UTP increased significantly. ATP levels increased approximately twofold in the first 6 h of germination. After 1 h without exogenous adenine, ATP dropped twofold or more in adenine auxotrophs but not in wild type. There was a net decrease in all adenine nucleotide pools during adenine starvation and a much greater decrease was seen in adenine auxotrophs than in wild type. The adenylate energy charge remained stable despite major changes in the adenylate pools.Accumulation of intermediates was observed in germinated conidia from purine auxotrophs blocked at various steps in the purine pathway. IMP accumulated in ungerminated and in starved conidia of the adenine-8 (ad-8) strain. Ungerminated conidia of the ad-5 strain contained a large pool of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) while the ad-1 strain had smaller amounts of AICAR, but significantly more than any other strain tested. The AICAR pools disappeared from ad-5 and ad-1 in the presence of 50 mg histidine/100 mL. Similarly the IMP pools in ad-8 decreased markedly in the presence of histidine, indicating that the contribution from the histidine biosynthetic pathway to purine nucleotide formation is significant.

Effect of cyclosporine on oxidative phosphorylation and adenylate energy charge of regenerating rat liver

1989 ◽  
Vol 189 (5) ◽  
pp. 313-320 ◽  
Author(s):  
S. Uemoto ◽  
K. Tanaka ◽  
K. Asonuma ◽  
R. Okamura ◽  
Y. Kitakado ◽  
...  
Keyword(s):  
Oxidative Phosphorylation ◽  
Rat Liver ◽  
Energy Charge ◽  
Adenylate Energy Charge ◽  
Regenerating Rat Liver

Changes in the adenylate energy charge ofNematospiroides dubiusandTrichostrongylus colubriformisparalysed by levamisolein vivo

Parasitology ◽  
1980 ◽  
Vol 81 (3) ◽  
pp. 593-601 ◽  
Author(s):  
M. J. Sharpe
Keyword(s):  
Oxygen Tension ◽  
Adenine Nucleotide ◽  
Energy Charge ◽  
Adenylate Energy Charge ◽  
Laboratory Mice ◽  
Trichostrongylus Colubriformis ◽  
Nucleotide Content ◽  
Nematospiroides Dubius ◽  
Low Levels

SUMMARYThe adenine nucleotide content and adenylate energy charge ofNematospiroides dubiusfrom laboratory mice and ofTrichostrongylus colubriformisfrom lambs has been measured. Administration of the anthelmintic, levamisole, to infected hosts resulted in only a slight fall in the adenylate energy charge ofN. dubiusover a 3-h period but there was a greater fall in the adenylate energy charge ofT. colubriformisduring this period. In neither case did the energy charge fall quickly, nor did it fall to the low levels which would be expected if the levamisole were inhibiting synthesis of ATP. The changes in energy charge of the nematodes which occurred following administration of levamisole to their hosts was of the order which can be satisfactorily explained by changes in the environment of the nematodes, such as reduced oxygen tension. It is concluded that the maintenance of levamisole-induced paralysis of these two species of trichostrongylein vivodoes not rely on the inhibition of fumarate reductase.

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